Method of producing nutritional products from human milk tissue and compositions thereof

ABSTRACT

Methods of isolating, treating, storing and processing human milk, as well as nutritional formulations of human milk comprising protective human milk proteins.

This application claims priority to U.S. Provisional Patent ApplicationSer. No. 60/290,823, filed May 14, 2001.

FIELD OF THE INVENTION

The present invention relates to biopharmaceutical and nutritionalproducts obtained from human milk and mammary gland secretions. Moreparticularly, the present invention relates to methods of isolating,storing, transferring, processing, packaging and deliveringpharmaceutical and nutritional formulations comprising cells and tissuesof human milk, fractionated human milk components and specificallyreunited components of human milk. One embodiment of the presentinvention relates to improving the nutrition of low birth weightinfants. Another embodiment of the present invention relates to a noveltype of immunoglobulin useful in treating disorders, such as, forexample, cancer, immune disorders, gastrointestinal disorders,nutritional disorders and metabolic disorders.

BACKGROUND OF THE INVENTION

The lack of a standardized source of human milk, available for researchpurposes, has seriously hampered scientific investigation of human milkas well as the majority of the nearly 4,000 unique, species-specificmilks. Accordingly, it would be desirable to provide a standardizedsource of human milk that can be modified to reflect the various stagesof lactation and various immune responses.

Although the presence of immunoglobulins in human milk has beenacknowledged for a number of years, the specific role and function ofeach of the human milk immunoglobulins and their subclasses has beenpoorly understood. For the patient of any age suffering from an immunedisorder, the worldwide gamma globulin shortage is impacting care tosuch a degree that some patients are unable to obtain treatment.Purified immunoglobulins from human milk hold the potential for asolution for this worldwide shortage. Accordingly, it would be desirableto provide formulations comprising higher levels of immunoglobulinsisolated from human milk, as well as methods to deliver theseformulations to patients.

It has been known for a long time by physicians, scientists andnutritionists that the best food or nutrition supplied to an infant isits own mother's milk, i.e., fresh human milk. Recent research hasindicated that “species-specific” milk plays a significant role indisease prevention and the severity of disease when the infant doesbecome ill. Until recently, the reasons behind the superiority ofspecies-specific milk were not well understood, nor were the variouscomponents and the roles they play in development and diseaseprevention. It is recognized, however, that many situations arisewherein the infant cannot obtain its mother's milk and as a result asuitable replacement is desired. Artificial baby milks, predominantlybased on cow's milk, have been prepared and used to nourish an infantbut there is increasing evidence that infants fed artificial baby milkssuffer long-term ill consequences. It has been suggested that theexposure of an infant to any foreign proteins, such as the bovineprotein, during the first few days of life will increase the infant'schance of becoming afflicted with juvenile diabetes. Other ill effectsinclude allergies, lowered immunity, gastrointestinal disorders,respiratory disease and other associated etiology. Although much efforthas been made to improve synthetic infant milk formulas, attempting tomake them more closely simulate mother's milk, the presence of livingorganisms and other “species-specific” cells that act in a way totrigger other disease preventing mechanisms in the infant, these effortshave proven futile.

According to Jenness and Sloan, human milk contains three major groupsof constituents that carry strong “species-specific” and“organ-specific” missions: (1) constituents specific to both organ andspecies, including proteins and lipids; (2) constituents specific toorgan but not species, including lactose; and (3) constituents specificto species but not to organ, including albumin and some immunoglobulins.

Human milk is not a uniform body fluid; instead, it is a secretion fromthe mammary gland of constantly changing composition. In nature, thecomposition of human milk changes not only from day to day, but alsothroughout the course of a single day. While the reasons and outcome ofthese changes are not fully understood, it is intuitive to believe thatthese changes benefit the species and that substantial advantages may begained for the infant who is provided an opportunity to reap thebenefits of a modified formulation of 100% human milk. Accordingly, itwould be desirable to provide formulations comprising human milkproteins as nutritional supplements and therapeutics for patients inneed of gamma globulin therapy.

SUMMARY OF THE INVENTION

Compositions containing 100% human milk proteins, including theso-called host resistance factors (HRF) of human milk, as well as othernutrients, living cells, and components are useful when employed toenhance and improve outcomes for babies and children who are not able toobtain human milk from their mothers (or cannot obtain enough mother'smilk or mother's milk in the formulation needed due to immunodeficiencyof the mother) as well as other patients (including adults) sufferingfrom immune disorders, nutritional disorders and other diseases anddietary challenges.

The sequential administration of many of the human milk constituentsprovides substantial value to the recipient because of metabolic andcatabolic processes. It is at the core of the present invention toutilize such human milk constituents, in their processed form, in such asequential fashion as to provoke the same type of chain reaction in thebody. With this concept, the pairing of the processed milk tissue withthe sequential and differentiated delivery methods, patients may enjoy anew type of preventative and therapeutic medicine. Because human milkimmunoglobulins are specifically targeted to many diseases of thenewborn, as well as the protective functions of the mucus membranes ofthe newborn's body, and contain higher levels of IgA, IgD, IgM and IgE,the term “panoglobulin” or “lactapanoglobulin” has been coined for thisnewly identified formulation. In addition to fighting immune disorderswith a human-milk origin panaglobulin, patients preparing for surgery,chemotherapy, radiation or other “currently accepted, but destructive”therapies, may enjoy preliminary therapies that may mitigate the illeffects of their upcoming procedure. In the same fashion, the constantlychanging nature of species-specific milk allows for the inclusion of themammary gland as a laboratory of sorts, seeking not to simply initiateand artificially replicate structures like antibodies and proteins, butinstead, to produce a bonafide human-produced fluid that can beisolated, processed and delivered for a highly targeted use againstdisease.

One embodiment of the present invention provides a nutritionalformulation of isolated human milk containing protective human milkproteins or host resistance factors of human milk suitable for infantconsumption which can be directly administered to an infant.

Another embodiment of the present invention provides a method ofisolating human milk comprising the steps of collecting a sample ofhuman milk from a donor in a collection device, storing the sample ofmilk obtained from the donor, and processing the milk sample byconducting a nutritional analysis on the milk sample; fortifying thesample with heat-resistant nutrients, pasteurizing the sample;fortifying the pasteurized sample with heat-sensitive nutrients andtesting the sample to ensure successful pasteurization.

Yet another embodiment of the present invention provides a system fordelivering human milk to an infant. The system contains a feeding tubetreated to minimize adherence of milk fat to the interior of the feedingtube; a heated sheath surrounding the feeding tube and an enteral pumpremovably mounted to a motorized platform.

DETAILED DESCRIPTION OF THE INVENTION

The present invention and the methods of obtaining and using the presentinvention will be described in detail after setting forth preliminarydefinitions.

Definitions

The following definitions are provided to facilitate understanding ofcertain terms used in the present invention.

As used herein, “human milk” means any stage of human milk productionincluding the production of breast secretions not associated withlactation. These stages include, but are not limited to, colostrums,transitional milk and mature milk.

As used herein, “species-specific milk” means any milk that would beprocessed or formulated to provide an advantage of any kind to its ownoffspring.

As used herein, “second-best species-specific milk” means any milk thatwould be processed or formulated to provide a “close second” to its ownspecies-specific milk resulting in better outcomes than using thestandard bovine or soy based milk replacer.

The present invention describes a method that includes multiple stepsand processes to harvest or isolate, store, transfer, process, packageand deliver a variety of pharmaceutical and nutritional formulationscontaining cells and tissues comprising 100% human milk tissue,fractionated human milk tissue components and specifically reunitedcompounds, as well as novel methods and procedures to affect levels ofsuch fractionated human milk tissue components, isolate them from rawhuman milk and deliver them through various methods including (but notlimited to) ingestion, inhalation, intra-nasal administration, eyedrops, ear drops, enema, douche, lavage, transdermally, rectally,intravenously, intramuscularly injection, direct injection, directtopical application, ng tube and jg tube.

Additionally, these formulations may be delivered through any of thesemethods, but when delivered, the present invention describes a sequenceof delivery by which certain components or compounds will catabolize tocreate optimum conditions for the sequential delivery of an additionalcompound. For instance, if the formulation is nutritionally focused, thepresent invention provides a formulation that is specifically deliveredin the morning, with a different formulation delivered in the afternoonand evening.

Little is understood at this time, as to why the formulation ofmammalian milk evolves throughout the day. The present invention isdirected to a method by which this differentiation is preferred andwould create an improved outcome for the patient. Additionally, by anymethod, there may be an advantage to the “priming” of the patient by thedelivery of certain processed human milk components, thereby eliciting aresponse in the patient's body that will improve outcomes when the nexttreatment in the sequence is followed. This sequential treatment conceptwould not be limited to the method of delivery. Instead, the presentinvention relates to the possibility that multiple delivery methods mayactually trigger multiple advantageous responses in the patient,increasing the patients' chance of an improved outcome by coaxing thepatient's system into active collaboration with the treatment method.This method simulates the natural processes of the mammalian immunesystem, which cannot be described as any one “silver bullet” but aseries of complex communications between multiple cell structures andthe offending pathogen.

The present invention relates to use of the disclosed methods andformulations from all mammalian species and is not limited to humanbeings. Additionally, the present invention encompasses all breastfluids as a potential source for harvesting milk and immune cells, asthe mammary gland is a lymphoid organ, capable of producingimmunoglobulins with or without accompanying lactation.

EXAMPLES The following examples are intended to illustrate variousembodiments of the present invention and are not to be construed aslimiting the scope of the invention. Example 1 Gamma GlobulinFormulations

At the center of this invention, is the intent to solve the worldwideshortage of gamma globulin. The current source of gamma globulin isblood serum, and specifically IgG from human blood. The presentinvention discloses a prophetic inclination, based upon a 15-year studyof human milk, that a new form of gammaglobulin referred to herein as“panaglobulin,” “mammaglobulin” or “lactopanaglobulin” may replace thecurrent gamma globulin. Because higher levels of IgA and IgM are presentin human milk and colostrums, and a more diverse form of IgG as well,panaglobulins may provide protection beyond the scope of current gammaglobulin therapy. Manipulation of the levels of immunoglobulins andtheir subclasses will result in formulations that are targeted atspecific diseases or organ systems, making it possible to attack diseaseusing nature's pharmaceutical laboratory, the mammary gland.Furthermore, milk donors who have weaned their babies or have initiatedlactation without pregnancy could feasibly become human labs, becomingexposed through any method to mild strains of disease and producing theappropriate antibody in their milk. Since the breast is reactive to newexposures of pathogens, an array of new immunities can be produced tocombat such diseases. Whether these types of donors could produce enoughmilk to become a primary source remains to be seen, but at least thesedonors could provide a human lab for biosynthesizing disease specificantibodies that could be replicated later using other methods.

Colostrum contains high levels of immunoglobulins, a vital defensemechanism that protects the newly born. sIgA provides immediateprotection to the infant by lining the gastrointestinal system andproviding a first defense against dangerous pathogens like E. coli andother devastating disease organisms. The invention disclosesconcentrated, processed sIgA for use as a prevention or therapeutic forgut disorders in patients of all ages. Potency levels will depend uponthe severity of the disease, the general health of the patient and thecost of the processing.

Colostrum also contains IgG1, G2, G3, G4, IgM, IgD and trace amounts ofother human origin immunoglobulins. All of these immunoglobulinsfunction in a myriad of ways, targeting specific organs and diseasestates. Because the mammary gland is a lymphoid organ, it is capable ofsynthesizing immunoglobulins, especially the four IgG subclasses, makingit possible to achieve a higher level of IgG subclasses in breast fluidthan is present in human blood serum. This capability of the mammarygland is not limited to lactation, with measurable quantities of IgGpresent in breast ductal fluid from non-lactating women. Expressingductal fluid may provide protective advantages to the donor,specifically the cleansing of the breast ductal system, as disclosed ina prior patent application by the inventor. This invention envisionsbreast fluid from non-lactating women as a potential source of humanimmunoglobulins. Current research cites a wide variety of volume andconstituents present in colostrums, transitional milk and mature milkbut little information exists for the constituents present in the breastductal fluid of non-lactating women. The present invention is directedto the ability to influence the volume and constituents of breast ductalfluid through dietary and pharmaceutical manipulation.

For nutritional and pharmaceutical applications, other valuable proteinscontained in human milk include alpha-lactalbumin, beta-lactoglobulin,lactoferrin, serum albumin, lysozyme, and other proteins as well. Humanmilk has a higher proportion of alpha-lactalbumin and the hostresistance factors or anti-microbial proteins of human milk, whichinclude lactoferrin, lysozyme and secretory IgA, and account for 75% ofthe protein in human colostrum as compared with 39% in mature humanmilk. Additional human milk cells that provide substantial diseaseresistance in the newly born include lymphocytes, macrophages, andsecretory IgA. Lactoferrin is present in relatively high amounts inhuman milk as is lysozyme and bifidus-stimulating factors. A majorobjective of this invention is to provide techniques and routines forimproving the diet and feeding of infants, particularlyvery-low-birth-weight infants. By varying the levels of many of thesespecies-specific milk constituents, the invention will result in amyriad of formulations specially suited to a wide variety of medicalconditions.

Example 2 Collection of Donor Milk

U.S. Pat. No. 4,772,262, which is hereby incorporated by reference inits entirety, is directed to technology for milk removal. As disclosedin that patent, milk yields increase due to the sensory stimulusprovided by the patented breast pump equipment. When milk yieldincreases, the formulation of milk including many of the valuableimmunoglobulins also increase along with living cells, such asmacrophages and lymphocytes. Lipids also increase and the mother's bodyresponds to the stimulus by producing higher levels of prolactin thatwill trigger continuing milk supply and the secretion of additionalnutrients into her milk.

Example 3 Storage of Donor Milk

Previous methods of collecting donor milk failed to recognize theimportance of stimulation to the mammary gland as well as collectionchambers designed for the anaerobic collection and transfer of donormilk. The invention describes such a method as part of its uniquecollection, storage and transfer system. Additionally, the preservationof milk components and nutrients is paramount to the success of theinvention wherein harvesting of milk cells specific to the species willresult in pharmaceutical and nutritional improvements in outcomes forthe newly born or immune compromised patient. For that reason, it isimportant that the container in which the donor milk is stored, willpreserve and protect these vital milk constituents from harm due toultraviolet light and other damaging light rays. A UV coating oradditive, applied to the collection bottle during the molding process orafterwards as an exterior coating or sheath will ensure that lightdegradation does not occur.

Finally, the design of the donor milk collection bottle should make iteasy to draw off a sample of the donor milk without compromising theintegrity of the milk sample. A proprietary design allows for atwist-turn valve to open and release a small amount of donor milkthrough a one-way valve into a test vial. The one-way valve prevents anybacteria or other pathogen from contaminating the milk sample.Additionally, a “tear down” design will allow frozen milk to beprocessed immediately, without the necessity of waiting for the milk tothaw. The tear down feature will provide an easy pull-tab that willstrip the container from the frozen block of donor milk. The pull-tabwill feature a tag on which a bar code is attached, so that during thetear down process, a “lot” numbering system will track the pooled milkback to their original donors.

Example 4 Transfer of Donor Milk

Novel designs for refrigerated transfer units utilizing alternativeforms of energy and equipped with temperature indicator recorders ensurethat the milk has been maintained under safe conditions. A programmablechip that records temperature variations as well as handling conditions(rough treatment can compromise milk quality by breaking cell walls),prevents the opening of the transfer case upon arrival at the processingplant. The milk is automatically rejected for quality issues andquarantined for further scrutiny. The transfer unit will contain aprogrammable chip that stores the contents, origin of contents, dateshipped, date received, lot numbers and any other information requiredfor quality control, regulatory or other reasons.

Example 5 Hospital Based Testing and Processing of “Mothers Own” Milk

The invention discloses a total quality control system that encompassesboth routine and novel procedures and tests. For mothers wishing toprovide their own milk for use specifically with their own baby, onsighttesting will be done at the hospital. Standard donor screening will bedone in accordance with current recommendations and accepted practices.In the present day, no routine testing is done in this case, andfrequently the lack of testing causes consternation and concern in thephysician with the end result being that babies are being routinelydeprived of their mothers' milk. Upon questioning the areas of concern,several neonatologists indicated a concern for the presence of streetdrugs, disease pathogens and contaminants. To answer this concern, theinvention includes a series of quick tests, designed to screen for thepresence of the most common pathogens, drugs and contaminants. In orderto provide the most efficient form of testing, a series of pumped milkis pooled, mixed and tested. A report is provided to the neonatologistand also placed in the infant's chart. The mothers' milk, intended forher own baby, is housed in the milk laboratory, under optimum storageconditions. Again, a temperature indicator on each container of milkensures that milk has not been exposed to adverse conditions that maycause degradation or contamination. The temperature indicator isattached to a disposable cap that covers the container. In the event ofadverse circumstances, the temperature indicator activates a lockingmechanism and the milk is quarantined until further analysis can bedone.

Example 6 Onsight (Hospital Based) Delivery Methods for Mothers Own Milk

Of special concern in high risk neonatal units, is the loss of milk fatthrough feeding tubes used to feed very-low-birth-weight, sick orpre-term infants. A special design for extruded tubing employs a methodduring the manufacturing process, that will eliminate the problem of fatsticking to the inside of the tubing. After extrusion, a heat treatmentis applied to the inside of the tubing, via a “pull-through” rod. Aheated element, coupled with an anti-static element, of sorts, eliminatethe static charge while smoothing the “tackiness” of the interior tubingwall. Coupled with a heated sheath, used during the tube feeding to keepthe flow of milk warm, the fat loss can be substantially decreased. Agentle rocking motion, created by a motorized platform on which theenteral pump sits, provides constant agitation and prevents the pumpedmilk from separating. Additional design features prevent the fat fromclinging to the inside of the enteral syringe, in which the pumped milkis contained. A Teflon coating, or alternatively a silicone interior bagor collapsible bag made from a food safe polyvinyl may create additionalsolutions to this problem. Techniques associated with sequential feedingmethods may also mitigate the problems associated with single feedmethod. By utilizing sequential feeds, the “foremilk” formulation(simulating the composition of the first milk a baby receives during adirect feeding from the breast), is administered. Low in fat, but highin volume, this feed usually takes more time than the higher fat“hindmilk” feed. The hindmilk feed can be then administered from a pushsyringe specially designed to conserve a large amount of the fat thatnormally would have stayed in the long tubing associated with theearlier feed.

Example 7 Plant Processing Methods

In the practices of this invention the human milk proteins, includingthe so-called host resistance factors (HRF) of human milk, are preparedby chemically fractionating the same using standard techniques, such asthe Cohn method, from pooled donor milk. This method will form the basisfor the extraction of the immunoglobulins for the ultimate purpose ofpurification and processing into nutritional, IV and injectable forms.The present invention discloses a completely closed system forprocessing.

Under this system, there is no opportunity for contamination. When thedonor milk is received at the processing center, a representative samplefrom each donor lot is tested and cultured. The remaining samples in thelot are transferred to the freezer to hold until the cultures are read.From the strip-down phase to the spray drying of the final product, allprocessing occurs within a sealed system. After the lot has been clearedfor processing, the frozen containers of milk are placed in an anaerobicchamber where the strip down of the bottle occurs. A filter preventsparticles of stripped down plastic bottles from entering the processingsystem. The frozen chunks of donor milk are thawed, using a slow,continuous heat with a mild churning action. Once thawed, a nutritionalanalysis is performed to determine specific nutritional levels of thepooled donor milk. Depending upon the desired human milk formulation,the system automatically adjusts the formulation, using validatedsources from human milk origin, if augmentation above the levels of thedonor milk is desired. Fortification at this point is limited tonutrients that are not adversely affected by heat. As the fortificationis being done, the milk is gently churned. The pasteurization processtakes place, again, in the same closed system, using the Holder Methodof 62.5 C for 20 minutes of 56C for 30 minutes. After pasteurization,the milk is cooled. Second stage fortification occurs at this point,with the addition of previously processed immunoglobulins, as well asselected, 100% screened human milk cells. After processing, finaltesting is done to determine that the pasteurization process has beensuccessful.

The formulations and methods of the present invention may be embodied inother specific forms without departing from the teachings or essentialcharacteristics of the invention. The described embodiments aretherefore to be considered in all respects as illustrative and notrestrictive. The scope of the present invention is defined in thefollowing claims, rather than the previous description, and all changesthat come within the meaning and range of equivalency of the claims aretherefore to be embraced therein.

1. A nutritional formulation of isolated human milk comprisingprotective human milk proteins of human milk suitable for infantcomposition, wherein the nutritional formulation can be directlyadministered to an infant.
 2. The nutritional formulation of claim 1further comprising human milk cells.
 3. The nutritional formulation ofclaim 2, wherein the human milk cells comprise lymphocytes andmacrophages.
 4. The nutritional formulation of claim 1, wherein thehuman milk proteins comprise alpha-lactalbumin, beta-lactoglobulin,lactoferrin, serum albumin, lysozyme and secretory IgA.
 5. A method ofisolating human milk comprising the steps of: (a) collecting a sample ofhuman milk from a donor in a collection device, wherein the collectiondevice comprises a computer chip capable of recording temperaturevariations and handling conditions of the collection device; (b) storingthe sample of human milk obtained from the donor; and (c) processing thesample of human milk obtained from the donor, wherein processingcomprises the steps of: (i) conducting a nutritional analysis on thesample of human milk; (ii) conducting a first fortification of thesample of human milk with heat-resistant nutrients; (iii) pasteurizingthe sample of first fortified human milk; (iv) conducting a secondfortification of the sample of human milk with heat-sensitive nutrients;and (v) testing the sample of fortified human milk to determine whetherthe pasteurization was successful.
 6. The method of claim 5, wherein thecomputer chip is further capable of recording the origin of the sample,contents of the collection device, volume of the collection device,shipping dates of the sample or lot number of the sample.
 7. The methodof claim 5, wherein the sample of human milk is frozen prior to theprocessing step.
 8. The method of claim 5, wherein the collection devicefurther comprises a twist-turn valve to open and release an amount ofmilk through a one-way valve into a test vial.
 9. A system fordelivering human milk to an infant comprising: (a) a feeding tube havingan inside and an outside, wherein the inside of the feeding tube istreated to prevent adherence of milk fat to the interior of the feedingtube; (b) a heated sheath in communication with the outside of thefeeding tube; and (c) an enteral pump removably mounted to a motorizedplatform, the enteral pump capable of transferring human milk throughthe feeding tube from a storage container to the infant.
 10. The systemof claim 9, wherein inside of the feeding tube is coated with silicone.